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Fig. 4 | Arthritis Research & Therapy

Fig. 4

From: IL13Rα1 protects against rheumatoid arthritis by combating the apoptotic resistance of fibroblast-like synoviocytes

Fig. 4

IL13Rα1 interacts and stabilizes DR5. a Whole lysates from CoCl2 or Tm challenged RA FLSs were IP with anti-IL13Rα1 or anti-DR5 and IB by anti-DR5 or anti-IL13Rα1. b After transfection with Lv-IL13Rα1 or Lv-Ctrl for 24 h, IL13Rα1 (green) and DR5 (red) in RA FLSs from three samples were detected by IF. c Western blot of DR5 protein in RA FLSs following transfection with Lv-IL13Rα1 relative to Lv-Ctrl. d With the exposure of CoCl2 or Tm, RA FLSs were transfected with siIL13Rα1/siCtrl for 24 h, DR5 protein was measured by Western blot, and band intensity given underneath gel images was measured using ImageJ software. e, f With the exposure of CHX plus Tm, total proteins from Lv-IL13Rα1/Lv-Ctrl-transfected or siIL13Rα1/siCtrl-transfected RA FLSs were subjected to Western blot for analyzing DR5 expression as indicated time points. Band intensity was measured using ImageJ software and presented as fold change compared as indicated. g Silencing efficiency of siRNA targeting DR5 was detected by Western blot, and these two siRNAs were combined at an equal concentration for the following experiments. RA FLSs (n = 3) harboring Lv-IL13Rα1 or Lv-Ctrl were transfected with siDR5 or siCtrl, and the apoptotic rate (h) and Bax or Bcl protein (i) were detected by Annexin V staining and western blot. Band intensities are quantified and the Bax/Bcl ratio is summarized into bar charts (right). Data are representative of three independent experiments from three different RA samples (n = 3) with similar results. ns, not significant vs the mean ± SD of Lv-Ctrl in the presence of siDR5; *p < 0.05 vs the mean ± SD of IL13Rα1 in the presence of siCtrl

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